Difference between revisions of "Part:BBa K2999003"
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===Usage and Biology=== | ===Usage and Biology=== | ||
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YAU-China expressed the biofilm-damaged system in 2019, using extracts from each part of the system for experiments. | YAU-China expressed the biofilm-damaged system in 2019, using extracts from each part of the system for experiments. | ||
After overnight induction of the control strain, two strains expressing single enzyme and two strains co-expressing two enzymes, four strains were collected by centrifugation and suspended in PBS buffer of equal volume. The supernatant was absorbed and added into the biofilm of Pseudomonas aeruginosa cultured on the pre-PVC plate. After incubation for a period of time, the PVC plate was cleaned, stained with crystal violet for 10 minutes, and the remaining biomass was treated with anhydrous ethanol. The coatings were washed out and OD570 was detected by enzyme labeling. | After overnight induction of the control strain, two strains expressing single enzyme and two strains co-expressing two enzymes, four strains were collected by centrifugation and suspended in PBS buffer of equal volume. The supernatant was absorbed and added into the biofilm of Pseudomonas aeruginosa cultured on the pre-PVC plate. After incubation for a period of time, the PVC plate was cleaned, stained with crystal violet for 10 minutes, and the remaining biomass was treated with anhydrous ethanol. The coatings were washed out and OD570 was detected by enzyme labeling. | ||
| + | [[File:T--YAU-China--iGEM2019-BBa -4.jpg|500px|thumb|none|Figure 1. Effects of PelA and PslG on Biofilm]] | ||
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| + | [[File:T--YAU-China--iGEM2019-BBa -3.png|500px|thumb|none|Figure 2. Crystal violet staining of biofilm treated with extracts from control group and three groups]] | ||
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Latest revision as of 11:48, 15 October 2019
ptac+RBS+PelA+RBS+PslG
Biofilm is a complex bacterial community embedded in the extracellular matrix composed of proteins, extracellular DNA (eDNA) and extracellular polysaccharides. Pseudomonas aeruginosa PAO1 has two types of biofilm, Psl and Pel. This part is mainly used for encoding polysaccharide hydrolases PslG and PelA, and destroying the structure of biofilm.
Usage and Biology
YAU-China expressed the biofilm-damaged system in 2019, using extracts from each part of the system for experiments. After overnight induction of the control strain, two strains expressing single enzyme and two strains co-expressing two enzymes, four strains were collected by centrifugation and suspended in PBS buffer of equal volume. The supernatant was absorbed and added into the biofilm of Pseudomonas aeruginosa cultured on the pre-PVC plate. After incubation for a period of time, the PVC plate was cleaned, stained with crystal violet for 10 minutes, and the remaining biomass was treated with anhydrous ethanol. The coatings were washed out and OD570 was detected by enzyme labeling.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NotI site found at 769
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2922
Illegal BglII site found at 3798
Illegal BamHI site found at 1905
Illegal BamHI site found at 2070
Illegal XhoI site found at 3802 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 686
Illegal NgoMIV site found at 710
Illegal NgoMIV site found at 731
Illegal NgoMIV site found at 832
Illegal NgoMIV site found at 1058
Illegal NgoMIV site found at 1766
Illegal NgoMIV site found at 1815 - 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1652
Illegal BsaI site found at 2278
Illegal BsaI.rc site found at 1853